Primer and method for detecting polymorphism of VKORC1 and CYP2C9 genes by adopting pyrosequencing method
A pyrosequencing method, CYP2C91075A technology, applied in the field of molecular biology testing, can solve the problems of difficult to achieve high-throughput detection, high equipment and technical requirements, slow detection speed, etc., to achieve short detection cycle and high specificity , the effect of simple operation
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Embodiment 1
[0057] A primer for detecting polymorphisms of VKORC1 and CYP2C9 genes by pyrosequencing, wherein the polymorphic site detected by VKORC1 is VKORC1-1639G>A, and the polymorphic site detected by CYP2C9 is CYP2C9430C>T or CYP2C9430C>T And / or CYP2C91075A>C, the primers include:
[0058] (1) The amplification primers for the VKORC1-1639G>A gene are:
[0059] Upstream primer: 5'-TGTTGGCCAGGCTTGTCTTAAACT-3'
[0060] Downstream primer: 5'-TGGGAAGTCAAGCAAGAGAAGACC-3'
[0061] The sequencing primer of VKORC1-1639G>A gene is: 5'-GCGTGAGCCACCGCA-3'
[0062] (2) The amplification primers for CYP2C9430C>T gene are:
[0063] Upstream primer: 5'-CTCATGACGCTGCGGAATTTT-3'
[0064] Downstream primer: 5'-CCACCCTTGGTTTTTCTCAACTC-3'
[0065] The sequencing primer of CYP2C9430C>T gene is: 5'-GGGCTTCCTCTTGAAC-3'
[0066] (3) The amplification primers for CYP2C91075A>C gene are:
[0067] Upstream primer: 5'-CCAGGAAGAGATTGAACGTGTGA-3'
[0068] Downstream primer: 5'-TTTGGGGACTTCGAAAACATG-3'
...
Embodiment 2
[0093] A primer for detecting polymorphisms of VKORC1 and CYP2C9 genes by pyrosequencing, wherein the polymorphic site detected by VKORC1 is VKORC1-1639G>A, and the polymorphic site detected by CYP2C9 is CYP2C9430C>T or CYP2C9430C>T And / or CYP2C91075A>C, the primers include:
[0094] (1) The amplification primers for the VKORC1-1639G>A gene are:
[0095] Upstream primer: 5'-TGTTGGCCAGGCTTGTCTTAAACT-3'
[0096] Downstream primer: 5'-TGGGAAGTCAAGCAAGAGAAGACC-3'
[0097] The sequencing primer of VKORC1-1639G>A gene is: 5'-GCGTGAGCCACCGCA-3'
[0098] (2) The amplification primers for CYP2C9430C>T gene are:
[0099] Upstream primer: 5'-CTCATGACGCTGCGGAATTTT-3'
[0100] Downstream primer: 5'-CCACCCTTGGTTTTTCTCAACTC-3'
[0101] The sequencing primer of CYP2C9430C>T gene is: 5'-GGGCTTCCTCTTGAAC-3'
[0102] (3) The amplification primers for CYP2C91075A>C gene are:
[0103] Upstream primer: 5'-CCAGGAAGAGATTGAACGTGTGA-3'
[0104] Downstream primer: 5'-TTTGGGGACTTCGAAAACATG-3'
...
Embodiment 3
[0131] A primer for detecting polymorphisms of VKORC1 and CYP2C9 genes by pyrosequencing, wherein the polymorphic site detected by VKORC1 is VKORC1-1639G>A, and the polymorphic site detected by CYP2C9 is CYP2C9430C>T or CYP2C9430C>T And / or CYP2C91075A>C, the primers include:
[0132] (1) The amplification primers for the VKORC1-1639G>A gene are:
[0133] Upstream primer: 5'-TGTTGGCCAGGCTTGTCTTAAACT-3'
[0134] Downstream primer: 5'-TGGGAAGTCAAGCAAGAGAAGACC-3'
[0135] The sequencing primer of VKORC1-1639G>A gene is: 5'-GCGTGAGCCACCGCA-3'
[0136] (2) The amplification primers for CYP2C9430C>T gene are:
[0137] Upstream primer: 5'-CTCATGACGCTGCGGAATTTT-3'
[0138] Downstream primer: 5'-CCACCCTTGGTTTTTCTCAACTC-3'
[0139] The sequencing primer of CYP2C9430C>T gene is: 5'-GGGCTTCCTCTTGAAC-3'
[0140] (3) The amplification primers for CYP2C91075A>C gene are:
[0141] Upstream primer: 5'-CCAGGAAGAGATTGAACGTGTGA-3'
[0142] Downstream primer: 5'-TTTGGGGACTTCGAAAACATG-3'
...
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